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Cited by 1 publication
(2 citation statements)
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“…PCR and editing assay. As described elsewhere (10), the product of reverse transcription was subjected to nested PCR amplification with Vent polymerase (New England Biolabs) and an Idaho Technology thermal cycler. One of the final pair of primers was labeled at the 5Ј end with biotin so that the product could be bound to superparamagnetic beads, denatured, and reconverted to doublestranded DNA with reverse transcriptase and a primer 5Ј labeled with 32 P. The product released from the beads by digestion with SalI was tested for susceptibility to further digestion with NcoI and NlaIII.…”
Section: Methodsmentioning
confidence: 99%
See 1 more Smart Citation
“…PCR and editing assay. As described elsewhere (10), the product of reverse transcription was subjected to nested PCR amplification with Vent polymerase (New England Biolabs) and an Idaho Technology thermal cycler. One of the final pair of primers was labeled at the 5Ј end with biotin so that the product could be bound to superparamagnetic beads, denatured, and reconverted to doublestranded DNA with reverse transcriptase and a primer 5Ј labeled with 32 P. The product released from the beads by digestion with SalI was tested for susceptibility to further digestion with NcoI and NlaIII.…”
Section: Methodsmentioning
confidence: 99%
“…A limitation of the nucleotide sequencing method described above was sensitivity; for example, to detect nucleotide changes less frequent than 10%, it would be necessary to sequence much more than 10 clones. To solve this problem, we made use of a more sensitive editing assay (10). PCR amplification of RNAs edited at 1011 and/or 1012 can be detected via the creation of sites for the restriction enzymes NcoI and NlaIII.…”
Section: Vol 69 1995mentioning
confidence: 99%